Objective To investigate the effect of lentivirus-mediated RNA interference(RNAi) on the expression of PKCγ mRNA and protein in rat neurons.Methods Primary cultured cortical neurons from SD rat (16 days of pregnency) embryos were randomly divided into 3 groups with 6 wells in each group:control group (group C),negative group(group NC)and RNAi group.Group C received no treatment.Each well in group NC was given negative lentivirus 3 × 105.Each well in group RNAi was given the recombinant lentiviral vector containing PKCγ shRNA(LV-PKC7 shRNA).The expression of PKCT mRNA and protein in rat neurons was detected by RT-PCR and Western blot 5 days later.The interference efficiency was ealculated.Results Compared with group NC,the expression of PKCγ mRNA and protein was down-regulatedin group RNAi(P<0.05),but no significant change Was found in group NC(P>0.05).The interference efficiency of gene and protein were 99.3%and 85.2%respectively.Conclusion Lentivirus-mediated RNAi can down restate the expression of PKC7 gene and protein in rat neurons.

OBJECTIVE: To investigate whether the reactive oxygen species (ROS) and mitochondrial ATP-sensitive potassium (mitoKATP) channels were involved in delayed neuroprotection induced by sevoflurane on tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) levels. METHODS: Eighty-four male SD rats weighing 250 approximately 280 g, undergoing thread embolism of the right middle cerebral artery occlusion (MCAO) to cause focal ischemia for 2 h and then undergoing 24 h reperfusion, were randomly divided into 7 groups (n=12, each): a sham group(S), an ischemia-reperfusion group (I/R), a sevoflurane preconditioning group (Sevo), a 2-mercaptopropionylglycine (ROS scavenger)+sevoflurane group (MPG+Sevo), a 5-hydroxydecanoate (a mitoK(ATP) blocker) + sevoflurane group (5-HD+Sevo), an MPG group, and a 5-HD group. The protein level of TNF-alpha and IL-1beta in the cerebral issue was detected by enzyme linked immunosorbent assay (ELISA) and the expression of mRNA was measured by semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: Apoptosis index (AI), the protein level and the mRNA expression of TNF-alpha and IL-1beta were significantly higher in the I/R group than those of Group S. Pre-administration of sevoflurane could inhibit the increase of the protein level and the expression of mRNA of TNF-alpha, and IL-1beta and attenuate the cerebral damage induced by ischemia-reperfusion. Neuroprotection of sevoflurane preconditioning was abolished by MPG and 5-HD. However, MPG and 5-HD alone had no effect. CONCLUSION Sevoflurane can produce delayed protection against cerebral ischemia-reperfusion injury by down-regulating TNF-alpha, IL-1beta protein, and mRNA expression.
Animals ; Hypoxia-Ischemia, Brain ; complications ; drug therapy ; Infarction, Middle Cerebral Artery ; complications ; drug therapy ; Interleukin-1beta ; genetics ; metabolism ; Ischemic Preconditioning ; methods ; KATP Channels ; metabolism ; Male ; Methyl Ethers ; pharmacology ; therapeutic use ; Neuroprotective Agents ; pharmacology ; therapeutic use ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reactive Oxygen Species ; metabolism ; Reperfusion Injury ; etiology ; prevention & control ; Sevoflurane ; Tumor Necrosis Factor-alpha ; genetics ; metabolism

OBJECTIVE: To observe the effect of intrathecal injection of nuclear factor-κB (NF-κB) inhibitor of pyrrolidine dithiocarbamate (PDTC) on pain sensitivity thresholds and the expression of spinal cord CX3C chemokine receptor 1 (CX3CR1) in monoarthritis (MA) model in rats. METHODS: Forty-eight Sprague-Dawley rats were randomly divided into 4 groups (12 each) after successful intrathecal catheterization: (1) sham operation with physiological saline group (the sham group); (2) MA with normal saline group (the MA group); (3) 10 μL 100 μmol/L PDTC before MA (the PDTC pre-treatment group); (4)MA before 10 μL 100 μmol/L PDTC (the PDTC post-treatment group). Normal saline or PDTC was injected 5 d after the intrathecal catheterization. Pain sensitivity thresholds were measured in the 4 groups before and after the intrathecal injection at different time points. Rat monoarthritis model was subsequently built by injecting complete Freund's adjuvant (CFA) into the left ankle joint of the rats. On day 3 after the intrathecal injection, expression of microglia in the L₅ spinal cord segment was observed by immunohistochemical method, and the lumbar segments L₄-L₅ of spinal cord were taken to perform RT-PCR to examine the expression of NF-κB mRNA and CX3CR1 mRNA. RESULTS: Compared with the MA group, the pain sensitivity thresholds in the sham group, the PDTC pre-treatment group and the PDTC post-treatment group at each time point after the intrathecal injection increased significantly (P<0.05), while microglia in the L₅ spinal cord segment decreased significantly (P<0.05) and expression of CX3CR1 mRNA and NF-κB mRNA in the lumbar segments L₄-L₅ of spinal cord decreased significantly (P<0.05). CONCLUSION The hyperalgesic effect of the CFA-induced model of monoarthritis can be relieved by intrathecal injection of NF-κB inhibitor PDTC. Its mechanism is possibly related to NF-κB signal pathway which is involved in the formation of inflammatory pain through regulating CX3CR1 expression.
Animals ; Arthritis ; chemically induced ; complications ; CX3C Chemokine Receptor 1 ; Injections, Spinal ; Male ; NF-kappa B ; metabolism ; Pain ; drug therapy ; etiology ; metabolism ; Pain Threshold ; drug effects ; Pyrrolidines ; administration & dosage ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Chemokine ; antagonists & inhibitors ; metabolism ; Spinal Cord ; metabolism ; Thiocarbamates ; administration & dosage

OBJECTIVE: To investigate the incidence of awareness during general anesthesia and analyze the risk factors in anesthetic practice and patient populations. METHODS: A total of 2 300 patients who underwent general anesthesia were included. Perioperative data and anesthetic drugs were collected prospectively. Patients were interviewed twice postoperatively with the same structured questionnaire. Each patient was classified into categories as no awareness, possible awareness, and awareness. RESULTS: Twenty-one patients (0.91%) definitely reported awareness, and another 205 (8.91%) reported possible awareness. Few of the patients with awareness required psychological intervention. ASA physical status III-IV and propofol maintenance were associated risk factors of awareness. CONCLUSION The incidence of intraoperative awareness is high in the clinical practice in major medical centers.
Adult ; Anesthesia, General ; adverse effects ; Anesthetics, Intravenous ; adverse effects ; Awareness ; China ; epidemiology ; Female ; Humans ; Incidence ; Intraoperative Complications ; epidemiology ; Male ; Middle Aged ; Propofol ; adverse effects ; Risk Factors

Objective:To observe the clinical parameters and short-term prognosis of Tibetan high-altitude area children with congenital heart disease undergoing surgery and anaesthesia in low-altitude area,and to investigate the perioperative safety of the treatment.Methods:From January,2016 to December,2016,14 children with congenital heart disease who underwent surgery were assigned into 2 groups (n=7 each):the high-altitude area group (X group,children from Tibetan Autonomous Region) and the low-altitude area group (H group,children from Hunan Province).Echocardiography data,perioperative hemodynamic changes,postoperative recovery,complication and perioperative serum N terminal pro B type natriuretic peptide (NT-proBNP) levels were recorded.Results:There were no significant differences in cardiac structure and function between the 2 groups,while the incidence of pulmonary hypertension in the X group was significantly higher than that in the H group (P＜0.05).There were no significant differences in perioperative hemodynamics between the 2 groups (P＞0.05),while the duration in ICU in the X group was longer than that in the H group and the serum NT-proBNP level in the X group was higher than that in the H group (P＜0.05).Conclusion:For children with congenital heart disease in Tibetan high-altitude area,undergoing surgery in low-altitude area contributes to a steady perioperative hemodynamics and helps to increase the perioperative safety.There may be a higher risk of postoperative cardiac dysfunction in Tibetan children than that in low-altitude area.